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[1]杨权,王月月,刘炎光,等.大豆子叶节遗传转化体系优化及抗逆基因AtNHX5的转化研究[J].大豆科学,2015,34(02):205-211.[doi:10.11861/j.issn.1000-9841.2015.02.0205]
　YANG Quan,WANG Yueyue,LIU Yan-guang,et al.Study on Optimization of Soybean Cotyledonary Node Genetic Transformation System and the Transformation of Resistance Gene AtNHX5[J].Soybean Science,2015,34(02):205-211.[doi:10.11861/j.issn.1000-9841.2015.02.0205]

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大豆子叶节遗传转化体系优化及抗逆基因AtNHX5的转化研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第34卷期数: 2015年02期页码: 205-211 栏目: 出版日期: 2015-04-25

Title:: Study on Optimization of Soybean Cotyledonary Node Genetic Transformation System and the Transformation of Resistance Gene AtNHX5

作者:: 杨权¹; 王月月¹; 刘炎光¹; 蒋春志²; 张孟臣²; 张洪霞³; 张洁¹ ; 王冬梅¹; 1河北农业大学生命科学学院, 河北保定 071001;?
2河北省农林科学院粮油作物研究所，河北石家庄 050000；
3中国科学院上海植物生理研究所，上海 200032

Author(s):: YANG Quan¹; WANG Yueyue¹; LIU Yan-guang¹; JIANG Chun-zhi²; ZHANG Meng-chen²; ZHANG Hong-xia³; ZHANG Jie¹; WANG Dong-mei¹; 1.College of Life Science, Agricultural University of Hebei, Baoding 071001,China;?
2. Grain and Oil Crop Research Institute, Academy of Agriculture and Forestry of Hebei Province, Shijiazhuang 050000, China;?
3. Shanghai Institute of Plant Physiology, Chinese Academy of Sciences, Shanghai 200032, China

关键词:: 大豆; 子叶节; AtNHX5基因; 遗传转化; 除草剂

Keywords:: Soybean; Cotyledonary node; AtNHX5; Genetic transformation; Glufosinate

分类号:: S565.1

DOI:: 10.11861/j.issn.1000-9841.2015.02.0205

文献标志码:: A

摘要:: 以河北省优良大豆品种冀豆15、五星2号和NF-58的子叶节为受体材料，利用农杆菌介导法进行遗传转化，探讨了影响农杆菌侵染后子叶节不定芽诱导的因素。结果表明：以萌发6 d、4℃处理24 h的子叶节为外植体，农杆菌侵染后经超声波处理30 s、共培养基中添加20 mg?L^-1硝酸银，能够提高子叶节丛生芽诱导率，转化植株经草铵膦筛选以及PCR检测，T₀代转化率达0.97%。利用该体系对大豆品种五星2号进行AtNHX5基因的遗传转化，获得3株T₀代RT-PCR检测阳性植株，且2-1号T₁代阳性植株检测具有一定耐盐性，初步证明获得了转AtNHX5基因的大豆新材料。

Abstract:: In this study,soybean cotyledonary node of Jidou 15, Wuxing 2 and NF-58 were used as materials. An efficient Agrobacterium-mediated gene transformation system based on the examinations of several factors influencing transformation efficiency was developed for soybean. The results indicated that the optimum transformation conditions were as follow: seeds germinated 6 days, 4℃ low temperature treatment, 20 mg?L^-1?anti-oxidant silver nitrate during co-cultivation, ultrasonic treatment for infected explants 30 s. In the above condition, PCR-positive rate was 0.97% Using the optimized system an stress-resistant gene AtNHX5 was transformed Using the optimized system, the gene was transformed into soybean Wuxing 2. After detected the transgenic plants resistant to glufosinate by PCR, the transformation efficiency was 0.23% The expression of AtNHX5 was assessed by RT-PCR analysis.One positive plant of T₁?generation was obtained by detection of PCR, which has salt-tolerance. It preliminarily demonstrated that the target gene was integrated into the soybean genome.

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备注/Memo

基金项目：转基因生物新品种培育重大专项（2011ZX08004-002-002，2014ZX0800402B-001）

第一作者简介：杨权（1986-），女，硕士，主要从事植物抗逆生理及遗传转化研究。E-mail:yangquan19860918@hotmail.com。

通讯作者：张洁（1974-），女，博士，副教授，主要从事植物抗逆机理研究。E-mail:zhangjiezhaolh@163com;王冬梅（1963-），女，博士，教授，博导，主要从事植物逆境分子细胞生物学领域的研究工作。E-mail:dongmeiwang63@126.com。

更新日期/Last Update: 2015-06-07